Figure 5.
Western Blotting Analysis for MECR and Lipoylated Protein Content of Cell Extracts from Several Affected Individuals’ Fibroblast Cell Lines
(A) Western blotting of whole-cell extract from cell lines of affected individuals 1–3 (family A, II:1; family B, II:2; family C, II:2) demonstrates reduced lipoylation and decreased level of MECR compared to four control fibroblast cell lines. DLAT-LA: lipoyated E2 subunit of human pyruvate dehydrogenase. DLST-LA: lipoylated E2 subunit of human α-ketoglutarate dehydrogenase. β-Actin: loading control.
(B) Western blotting of whole-cell extract from cell lines of affected individual 6 (family E, II:1) shows decreased level of MECR compared to control fibroblast cell lines.
(C and D) Densitometric quantitation of the DLAT-LA (C) and DLST-LA (D) signals show significantly reduced lipoylation in affected individuals 1–3 fibroblasts compared with controls. For the DLAT/DLST control intensity bars, the average intensity of the luminescence signal of the four samples from healthy controls was calculated. The mean value was defined as 1.0 (100% intensity) and the error bars represent the standard error of the mean (SEM).